Related Experiment Video
Updated: Dec 25, 2025
Studying Interactions of Staphylococcus aureus with Neutrophils by Flow Cytometry and Time Lapse Microscopy
Published on: July 17, 2013
Detection of methicillin-resistant Staphylococcus aureus persistence in osteoblasts using imaging flow cytometry
Dafne Bongiorno1, Nicolò Musso2, Lorenzo Mattia Lazzaro1
1Department of Biomedical and Biotechnological Sciences (BIOMETEC), Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), University of Catania, Catania, Italy.
Methicillin-resistant Staphylococcus aureus (MRSA) clones show varied intracellular persistence in human osteoblasts. Imaging flow cytometry revealed significant differences in bacterial survival, highlighting clone-specific host-pathogen interactions crucial for chronic infections.
Area of Science:
- Microbiology
- Infectious Diseases
- Cell Biology
Background:
- Methicillin-resistant Staphylococcus aureus (MRSA) is a primary cause of persistent infections, including osteomyelitis and prosthetic joint infections.
- Understanding host-pathogen interactions is critical for developing effective treatments against MRSA.
- Specific MRSA strains, particularly those belonging to major ST-MRSA-SCCmec clones, exhibit distinct behaviors within host cells.
Purpose of the Study:
- To investigate and compare the internalization and intracellular persistence of well-characterized MRSA strains within human osteoblasts.
- To evaluate the efficacy of imaging flow cytometry (IFC) as an improved method for studying host-pathogen dynamics compared to traditional cell culture assays.
Main Methods:
- Human MG-63 osteoblasts were infected with various MRSA clones.
- Internalization was assessed at 3 hours post-infection.
- Intracellular bacterial persistence was evaluated at 24 hours post-infection using both routine cell culture and vancomycin-BODIPY stained IFC.
- Statistical analysis was performed to compare persistence rates among different MRSA clones.
Main Results:
- All tested MRSA clones demonstrated significant internalization into osteoblasts within 3 hours.
- Intracellular persistence varied significantly among MRSA clones at 24 hours, with ST239-SCCmecIII (47.82%) and ST22-SCCmecIVh (50.55%) showing higher rates.
- IFC provided greater reproducibility and allowed analysis of live host cells, offering advantages over standard cell culture methods.
- The study identified heterogeneity in the persistence capabilities of different HA-MRSA clones within host cells.
Conclusions:
- MRSA clones exhibit distinct intracellular persistence behaviors within human osteoblasts, influenced by the specific clone and the number of infected cells.
- Imaging flow cytometry is a robust and advantageous technique for quantifying intracellular bacterial persistence.
- These findings underscore the heterogeneity of MRSA clones and their potential to cause persistent infections, informing future therapeutic strategies.
